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Methods for sequencing of partial ribose desoxyribonucleic acid (rDNA) of mitochondria 12s genes of brachionus rotifer and identifying brachionus rotifer

A technology of mitochondria and rotifers, applied in the biological field

Inactive Publication Date: 2014-06-04
ANHUI NORMAL UNIV
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Problems solved by technology

The identification of rotifers by molecular biology solves the identification difficulties of using morphology and ultrastructure, so that in the process of large-scale cultivation of rotifers, the population competition caused by population mixing can be reduced, resulting in reduced production of rotifers or even possibility of mass death

Method used

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Embodiment Construction

[0014] The biological materials used in this method for the identification of rotifers of the genus Brachionus are Brachionus rotifers, Brachionus calyxae, Brachionus acanthus, Brachionus square, Brachionus schizopoda, and Sickle rotifers. Brachionus rotifers, Brachionus tendigitata, Brachionus occipitalis, and Brachionus citriformum, pick a single individual of each rotifer, and culture them in monoclonal manner, among which Brachionus redus and Brachionus square , Brachionus citriformum, and Brachionus angularis were formulated by Gilbert (1963), and the other species used the original local water samples as culture medium after filtration. The temperature was basically consistent with the ambient temperature when they were collected. The food used was HB -4 culture medium, Scenedesmus obliquus in the exponential growth phase, when the number of single clone rotifers reaches a certain amount, filter the rotifers with a 25 μm pore size filter, and rinse with sterile distilled ...

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Abstract

The invention relates to methods for sequencing of partial ribose desoxyribonucleic acid (rDNA) of mitochondria ND12s genes of a brachionus rotifer and identifying the brachionus rotifer. A sequencing method comprises total DNA extraction of the brachionus rotifer, polymerase chain reaction (PCR) primer amplification and sequence determination, and phylogeny analysis. An identifying method uses the mitochondria ND12s rDNA of the brachionus rotifer to carry out identification. PCR primer designed in the sequencing method can amplify the mitochondria ND12s rDNA of the brachionus rotifer and others, a sequencing result can be used for accurate identification for brachionus rubens and others, the problems of morphology utilization and an identification difficulty existing in an ultrastructure are resolved, and the probability of rotifer production reduction or even mass mortality caused by population competition brought by population mingle is reduced in the process of large-scale rotifer cultivation.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a partial rDNA sequencing of the mitochondrial 12s gene of the rotifer of the genus Brachion and a method for identifying the rotifer of the genus Brachiocauda. Background technique [0002] Early taxonomic and systematic studies of rotifers were mainly based on morphological data. For example, Wallace (1989) conducted cluster analysis using eight morphological features and found that Rotifera, Eutatoria, and Monogonota were monophyletic, and clustered using nine morphological features. Class analysis found that the genus Notholca was divided into six clades. Melone (1998) obtained a parsimony clustering tree through matrix analysis of 60 morphological features of 1 acanthocephala, 6 rotifers and 2 outgroups (planarians and gnathogastrics), and found that the Dinestidate ( Digononta) is a paraphyletic group, but the monophyletic of rotifers cannot be determined....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 程双怀张宁张逸夏梦宁刘应龙卢燕梅闫锦锦
Owner ANHUI NORMAL UNIV
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